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ObjectiveTo observe the effect of Scutellariae Radix-Coptidis Rhizoma on plaque stability in atherosclerotic (AS) mice and to explore its possible mechanism of action based on the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa B (NF-κB) signaling pathway. MethodTen normal C57BL/6J mice were used as the normal group, and the same strain of ApoE knockout (ApoE-/-) mice were fed with a high-fat diet for 12 weeks to construct an atherosclerosis model. Mice were randomly divided into five groups, namely the model group, the atorvastatin group, and the Scutellariae Radix-Coptidis Rhizoma low-, medium-, and high-dose groups, with ten mice in each group. Then normal and model groups were given equal volume of saline gavage, and the low-, medium-, high-dose Scutellariae Radix-Coptidis Rhizoma groups were given 1.95, 3.9, 7.8 g·kg-1 of the drug by gavage for 8 weeks, respectively. The general state of mice was observed. Hematoxylin-eosin staining was utilized to observe the pathology of aortic root plaques and calculate the percentage of plaque area. Masson staining and oil red O staining combined with immunohistochemistry of F4/80 and α-SMA were used to detect the plaque components of aortic root plaques and calculate the plaque vulnerability index. Enzyme-linked immunosorbent assay was adopted to detect the expression levels of serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). Western blot was applied to detect the protein expression levels of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), TLR4, MyD88, NF-κB p65, and phosphorylation (p) -NF-κB p65 in the aortic tissues of mice in each group. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) assay was employed to detect the expression levels of MMP-2, MMP-9, TLR4, and MyD88, NF-κB p65 mRNA. ResultCompared with the model group, the general state of the mice in each medication group was improved, and no obvious side effects were observed. Compared with the model group, the percentage of plaque area in the aortic root of AS mice was significantly reduced in the medium- and high-dose Scutellariae Radix-Coptidis Rhizoma groups (P<0.05). The content of collagen fibers and smooth muscle cells in the plaques of the high-dose Scutellariae Radix-Coptidis Rhizoma group was significantly increased (P<0.01), and the content of lipids and macrophages was significantly reduced (P<0.05), the plaque vulnerability index of each dose group of Scutellariae Radix-Coptidis Rhizoma was significantly reduced, with significant reduction of the medium- and high-dose groups (P<0.01). MMP-2 and MMP-9 protein and mRNA expression levels in aortic tissues were significantly reduced in medium- and high-dose Scutellariae Radix-Coptidis Rhizoma groups (P<0.05). The serum levels of TNF-α and IL-6 were significantly reduced in AS mice in medium- and high-dose Scutellariae Radix-Coptidis Rhizoma groups (P<0.05). In the medium- and high-dose Scutellariae Radix-Coptidis Rhizoma groups, the levels of TLR4, MyD88 protein, and mRNA expression in aortic tissues were significantly reduced (P<0.05), and the level of NF-κB p65 phosphorylation in aortic tissues was significantly reduced (P<0.05). ConclusionScutellariae Radix-Coptidis Rhizoma may play an anti-inflammatory and stabilizing role by inhibiting the TLR4/MyD88/NF-κB signaling pathway.
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ObjectiveTo investigate the therapeutic effect of Scutellariae Radix-Coptidis Rhizoma (SRCR) on atherosclerosis (AS) in mice and the effect of SRCR on macrophage pyroptosis in plaques via NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasomes. MethodApoE-/- mice were fed with a high-fat diet for the modeling of AS and randomized into model, atorvastatin (5 mg·kg-1), and low-, medium-, and high-dose (1.95, 3.9, 7.8 g·kg-1, respectively) SRCR groups. Normal C57BL/6J mice were selected as the control group. After 8 weeks of administration, hematoxylin-eosin staining was used to observe the pathological status of the aortic plaque. The lipid accumulation in aortic plaque was observed by oil red O staining. The serum levels of total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) in mice were measured. Immunofluorescence double staining was employed to detect the co-localized expression of EGF-like module-containing mucin-like hormone receptor-like 1 (EMR1)/NLRP3 and EMR1/gasdermin D (GSDMD). The serum levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) were determined by enzyme-linked immunosorbent assay (ELISA). The protein levels of NLRP3, apoptosis-associated speck-like protein (ASC), Caspase-1, cleaved Caspase-1, GSDMD, N-terminus of GSDMD (GSDMD-NT), pro-IL-1β, IL-1β, and IL-18 were determined by Western blot, and the mRNA levels of NLRP3, ASC, Caspase-1, GSDMD, IL-1β, and IL-18 were determined by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the control group, the model group showed obvious plaques, elevated serum levels of TG, TC, LDL-C, IL-1β, and IL-18 (P<0.01), lowered serum level of HDL-C (P<0.01), and up-regulated expression of NLRP3 inflammasomes and molecules related to pyroptosis in the aortic plaques (P<0.01). Compared with the model group, SRCR, especially at the medium and high doses, alleviated the plaque pathology, reduced the lipid content in plaques (P<0.05, P<0.01), recovered the serum lipid levels (P<0.05), reduced the macrophage recruitment (P<0.01), activation of NLRP3 inflammasomes, and pyroptosis in aortic root plaques (P<0.05), lowered the serum IL-1β and IL-18 levels (P<0.01), and down-regulated the protein levels of NLRP3, ASC, Caspase-1, cleaved Caspase-1, GSDMD, GSDMD-NT, pro-IL-1β, IL-1β, and IL-18 (P<0.05) and the mRNA levels of NLRP3, ASC, Caspase-1, GSDMD, IL-1β, and IL-18 in the aortic tissue (P<0.05). ConclusionSRCR exerts a therapeutic effect on high-fat diet-induced AS in mice by inhibiting the activation NLRP3 inflammasomes and reducing the pyroptosis of macrophages in plaques.
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By consulting ancient herbal books and modern literature, this paper systematically sorted out and researched the processing history, relevant processing norms in recent years, modern processing technology, chemical composition changes of processed products and their pharmacological mechanism of Scutellariae Radix, in order to provide a basis for the further development of Scutellariae Radix decoction pieces. According to the textual research of ancient books, there were many kinds of processing auxiliary materials of Scutellariae Radix, such as wine, vinegar, salt, honey, pig bile and so on, among which the wine processing was the most diverse and detailed, and the processed products such as raw products, stir-fried products, wine-processed products, fried charcoal products were still in use. The modern processing techniques of Scutellariae Radix mainly focus on the processing aspects of softening and slicing, wine processing and charcoal frying, and the research methods are relatively unified. At present, it is found that the changed chemical constituents of Scutellariae Radix after processing are flavonoids, polysaccharides, volatile oils and trace elements, etc. Pharmacological effects of processed products are hemostasis, antibacterial, anti-inflammatory, antioxidant, analgesic and antipyretic, treatment of lung diseases, treatment of colitis, etc. However, in the studies of Scutellariae Radix processing, there is a lack of research on the structural changes of chemical components caused by processing and a comprehensive comparative study on the pharmacological effects of various processed products. Based on this, it is suggested to carry out systematic research on the processing technology to processing mechanism, further explore the relationship between the change rule of material basis and pharmacological action before and after processing of Scutellariae Radix, and deepen the exploration of molecular mechanism and clinical application of processed products of Scutellariae Radix, in order to clarify the scientific connotation of the processing mechanism of Scutellariae Radix, and lay a foundation for the subsequent expansion of the application of Scutellariae Radix decoction pieces and the formulation of processing standards.
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Scutellariae Radix-Coptidis Rhizoma(SR-CR) herbal pair is commonly used in many compound prescriptions for their synergistic heat-clearing and dampness-drying properties. During the decoction process, a substantial amount of precipitate is generated. However, there have been no explicit reports on the composition, morphology, and potential effects of this precipitate on the in vivo behavior of SR-CR decoction. This study employed high-performance liquid chromatography(HPLC), high-resolution mass spectrometry, and other techniques to analyze the composition of the co-precipitate in the decoction of SR-CR. Scanning electron microscopy and mass spectrometry imaging were used to analyze its appearance and morphology. Additionally, rats were used to investigate the effects of the co-precipitate on the in vivo behavior of the main components in the SR-CR decoction. The research findings indicated that eight components, including coptisine, berberine, epiberberine, palmatine, baicalin, oroxylin A-7-O-β-D-glucuronide, wogonoside and baicalein, constituted the primary composition of the co-precipitate. Among these, baicalin and berberine hydrochloride were the most abundant, accounting for about 60% of the total weight. Moreover, the co-precipitate contained 18% tannins. Morphological analysis revealed that the particles in the SR-CR decoction precipitate were spherical microparticles with an average diameter of around 600 nm. Pharmacokinetic research demonstrated that there were significant differences in the AUC, C_(max), t_(1/2), and T_(max) of baicalin, a major component, in rats administered with lyophilized powders of the combined decoction and single decoctions of SR-CR orally, suggesting that the precipitate generated during the decoction process can affect the in vivo behavior of the main components of the SR-CR decoction. It can reduce the absorption of baicalin in the body, decrease the extent of rapid drug release, and to a certain extent, prevent adverse reactions or side effects.
Subject(s)
Rats , Animals , Drugs, Chinese Herbal/pharmacology , Scutellaria baicalensis/chemistry , Berberine , Chromatography, High Pressure Liquid , Mass SpectrometryABSTRACT
This study investigated the mechanism of action of Scutellariae Radix-Coptidis Rhizoma(SR-CR) in intervening in non-alcoholic fatty liver disease(NAFLD) in rats based on lipidomics. Thirty-six SD rats were divided into a control group, a model group, SR-CR groups of different doses, and a simvastatin group, with six rats in each group. Rats in the control group were fed on a normal diet, while those in the remaining groups were fed on a high-lipid diet. After four weeks of feeding, drug treatment was carried out and rats were sacrificed after 12 weeks. Serum liver function and lipid indexes were detected using kits, and the pathomorphology of liver tissues was evaluated by hematoxylin-eosin(HE) staining and oil red O staining. Changes in lipid levels in rats were detected using the LC-MS technique. Differential lipid metabolites were screened by multivariate statistical analysis, and lipid metabolic pathways were plotted. The changes in lipid-related protein levels were further verified by Western blot. The results showed that compared with the control group, the model group showed increased levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), total cholesterol(TC), triglyceride(TG), and low-density lipoprotein cholesterol(LDL-c)(P<0.01), and decreased levels of γ-glutamyl transferase(γ-GT) and high-density lipoprotein cholesterol(HDL-c)(P<0.01), which were significantly recovered by the intervention of SR-CR. HE staining and oil red O staining showed that different doses of SR-CR could reverse the steatosis in the rat liver in a dose-dependent manner. After lipidomics analysis, there were significant differences in lipid metabolism between the model group and the control group, with 54 lipids significantly altered, mainly including glycerolipids, phosphatidylcholine, and sphingolipids. After administration, 44 differential lipids tended to normal levels, which indicated that SR-CR groups of different doses significantly improved the lipid metabolism level in NAFLD rats. Western blot showed that SR-CR significantly decreased TG-synthesis enzyme 1(DGAT1), recombinant lipin 1(LPIN1), fatty acid synthase(FASN), acetyl-CoA carboxylase 1(ACC1), and increased the phosphorylation level of ACC1. These changes significantly decreased the synthesis of TG and increased the rate of its decomposition, which enhanced the level of lipid metabolism in the body and finally achieved the lipid-lowering effect. SR-CR can improve NAFLD by inhibiting the synthesis of fatty acids and TG.
Subject(s)
Rats , Animals , Non-alcoholic Fatty Liver Disease/drug therapy , Scutellaria baicalensis , Drugs, Chinese Herbal/therapeutic use , Pharmaceutical Preparations , Rats, Sprague-Dawley , Liver , Triglycerides/metabolism , Cholesterol , Diet, High-Fat , Azo CompoundsABSTRACT
Objective:To investigate the changes of chemical constituents of Scutellariae Radix before and after carbonizing.Methods:Fifteen batches of Scutellariae Radix were collected. Diamonsil C18 (250 mm×4.6 mm, 5 μm) column was used; column temperature was 30 ℃; sample size was 10 μl and detection wavelength was 274 nm. The mobile phase A was 1% formic acid aqueous and B methanol, and the flow rate was 1 ml/min. The fingerprint of Scutellariae Radix before and after carbonizing was established and the changes of related components were analyzed. The content of tannin before and after processing was determined by phosphomolybdenum-tungstic acid-casein colorimetric method with gallic acid as the control.Results:Baicalin, baicalein, wogonoside and wogonin were identified by HPLC fingerprint. The contents of baicalin and wogonoside decreased, while the contents of baicalein and wogonin increased. The tannin content of Scutellariae Radix was significantly reduced after stir-frying.Conclusion:The glycosides are first converted into aglycones, then the aglycones are destroyed and the tannins are significantly reduced, which could provide a reference for the quality control of carbonized Scutellariae Radix.
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Dead heart is an important trait of pith-decayed Scutellariae Radix. The purpose of this study was to clarify the scientific connotation of the dead heart using multi-omics. Metabolomics and transcriptomics combined with multivariate statistical analysis such as principal component analysis(PCA) and partial least squares discriminant analysis(PLS-DA) were used to systematically compare the differences in chemical composition and gene expression among phloem, outer xylem and near-dead xylem of pith-decayed Scutella-riae Radix. The results revealed significant differences in the contents of flavonoid glycosides and aglycones among the three parts. Compared with phloem and outer xylem, near-dead xylem had markedly lowered content of flavonoid glycosides(including baicalin, norwogonin-7-O-β-D-glucuronide, oroxylin A-7-O-β-D-glucuronide, and wogonoside) while markedly increased content of aglycones(including 3,5,7,2',6'-pentahydroxy dihydroflavone, baicalin, wogonin, and oroxylin A). The differentially expressed genes were mainly concentrated in KEGG pathways such as phenylpropanoid metabolism, flavonoid biosynthesis, ABC transporter, and plant MAPK signal transduction pathway. This study systematically elucidated the material basis of the dead heart of pith-decayed Scutellariae Radix with multiple growing years. Specifically, the content of flavonoid aglycones was significantly increased in the near-dead xylem, and the gene expression of metabolic pathways such as flavonoid glycoside hydrolysis, interxylary cork development and programmed apoptosis was significantly up-regulated. This study provided a theoretical basis for guiding the high-quality production of pith-decayed Scutellariae Radix.
Subject(s)
Drugs, Chinese Herbal/chemistry , Scutellaria baicalensis/chemistry , Glucuronides , Multiomics , Flavonoids/chemistryABSTRACT
Scutellariae Radix, Coptidis Rhizoma, and Phellodendri Chinensis Cortex are three commonly used Chinese herbal medicines, and their application in classic famous prescriptions cannot be fully explained by the triple energizer classification method. This study reviewed the ancient materia medica works and elaborated on the functions of the three herbal medicines before the Song dynasty and after the Jin dynasty. The works before the Song dynasty mainly introduced the diseases treated by the three herbal medicines according to the Shennong's Classic of Materia Medica(《神农本草经》), and the works after the Jin dynasty mainly expounded the indications of the three medicines in the Shennong's Classic of Materia Medica according to the new medical theory. Although all the three herbal medicines can treat heat syndrome, digestive system diseases, skin and mucosa diseases, they act on different targets. Scutellariae Radix can regulate Qi stagnation and reverse caused by the fire syndrome. Coptidis Rhizoma can treat excess fire and purulent bloody stool caused by the deficiency of Zang-fu organs. Phellodendri Chinensis Cortex mainly treats the diseases of the intestine and reproductive system and can kill parasites. In addition, this paper summarized the descriptions about the functions of Scutellariae Radix in eliminating blood blockage, Coptidis Rhizoma in protecting the intestine, and Phellodendri Chinensis Cortex in clearing the liver in ancient books. According to the sentences in the Treatise on Febrile and Miscellaneous Diseases(《伤寒杂病论》), the application of Scutellariae Radix in Chaihu prescriptions, Coptidis Rhizoma in Baitouweng Decoction, and Phellodendri Chinensis Cortex in Zhizi Baipi Decoction confirms to the indications of the three medicines in the materia medica works before the Song Dynasty. The existing clinical and pharmacological research results confirm the indications of the three herbal medicines in the ancient works. The clinical application of the three medicines should refer to the materia medicia works before the Song dynasty, so as to achieve precise medication.
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By reviewing ancient materia medica, prescription books, medical books and modern literature, this article makes a systematic textual research on the name, origin, commercial specification, producing area, harvesting and processing of Scutellariae Radix used in famous classical formulas according to the historical development, in order to provide a reference for the development and utilization of famous classical formulas. Scutellariae Radix was first named as Qian, Qin and so on, while Huangqin was used as the official name in ancient literature, as well as many aliases such as Fuchang, Huangwen and Dufu. The main origin of Scutellariae Radix in the past dynasties was Scutellaria baicalensis, and other Scutellaria plants were also used for Scutellariae Radix at different times. The medicinal parts of Scutellariae Radix in all dynasties are roots, since the Northern and Southern dynasties, it has gradually differentiated into Ziqin and Kuqin with different efficacy according to their different growth stages and characteristics, and continued to this day. At present, most of Scutellariae Radix are Ziqin in the market, due to the influence of cultivation cost, index component content requirements and other factors. The production area of Scutellariae Radix recorded in ancient literature was wide and gradually expanded from the Yangtze River basin to the northward. Since modern times, Chengde city of Hebei province has been respected as a geo-authentic area, now S. baicalensis is cultivated in a wide area, spreading over north and northeast China. Scutellariae Radix was mostly used as raw products in the early period, and a variety of processed products appeared in the Ming dynasty. Development continues to this day, Scutellariae Radix mainly includes raw products and wine-processed products. According to the research conclusion, it is suggested that S. baicalensis should be used as Scutellariae Radix in famous classical formulas, and the selection of its commercial specifications and processed products can be comprehensively determined according to the requirements of original prescription and the clinical effect.
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ObjectiveTo observe the effects of Scutellariae Radix (SR)-Paeoniae Radix Rubra (PRR) combination of different proportions on the expression of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor κB (NF-κB) and phosphatidylinositol kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways in liver tissues of rats with hepatic fibrosis and explore the mechanism against hepatic fibrosis. MethodSixty male SD rats of SPF grade were randomly divided into a normal group, a model group, a positive control (silymarin) group, and SR-PRR 1∶1, SR-PRR 1∶2, and SR-PRR 1∶4 groups, with 10 rats in each group. The hepatic fibrosis model was induced in rats except for those in the normal group by intraperitoneal injection of 40% tetrachloromethane (CCl4)-olive oil solution at 3 mL·kg-1, 5 mL·kg-1 for the first time, for 8 weeks, twice per week. After 4 weeks, rats were treated correspondingly at 10 mL·kg-1 by intragastric administration, and the body weight of rats in each group was weighed for 8 weeks. After administration, histopathological changes in the liver were observed. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), albumin (ALB), alkaline phosphatase (AKP), and superoxide dismutase (SOD) activities, malondialdehyde (MDA), and hydroxyproline (HYP) content in liver tissues were detected. The mRNA expression levels of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR). ResultCompared with the model group, SR-PRR combination of different proportions could recover the body weight and improve the pathological injury of the liver. As revealed by enzyme linked immunosorbent assay (ELISA) results, compared with the normal group, the model group showed increased ALT, AST, HA, LN, AKP, MDA, and HYP levels to different degrees (P<0.05). Compared with the model group, the groups with drug intervention showed decreased levels of ALT, AST, HA, LN, AKP, MDA, and HYP, potentiated SOD activity, and increased level of ALB (P<0.05). As revealed by Real-time PCR results, compared with the normal group, the model group showed increased mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR (P<0.05). Compared with the model group, the groups with drug intervention showed reduced mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats (P<0.05). ConclusionSR-PRR combination of different proportions can improve the histopathological injury in liver tissues caused by CCl4, with the optimal effect observed in the SR-PRR 1∶4 group. SR-PRR may inhibit the development of liver fibrosis by inhibiting the expression of TLR4/MyD88/NF-κB and PI3K/Akt/mTOR signaling pathways, thereby alleviating chemical-induced liver injury.
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ObjectiveTo obtain content characteristics of inorganic elements in Scutellariae Radix (aged 1-4 years), and to explore the feasibility of identifying the growth years of Scutellariae Radix based on characteristic spectrum of inorganic elements combined with chemometric models. MethodAfter microwave digestion, the contents of Mn, Zn, Ca, Fe, Mg, Na, K, Cr, Cu, Se, As, Cd, Hg, Pb and Ni in 21 batches of Scutellariae Radix were determined by inductively coupled plasma atomic emission spectrometry (ICP-OES) and inductively coupled plasma mass spectrometry (ICP-MS). Meanwhile, characteristic spectrum of inorganic elements in samples was drawn. The identification model was constructed to discriminate the growth years of Scutellariae Radix based on the combination of principal component analysis (PCA), Fisher discriminant function and support vector machine (SVM). ResultThe contents of Mn (7.79-36.48 μg·g-1), Zn (10.12-31.43 μg·g-1), Cu (6.38-17.20 μg·g-1), K (2.98-13.89 μg·g-1), Mg (3.45-7.78 μg·g-1) and Ca (2.32-7.09 μg·g-1) in Scutellariae Radix were detected by ICP-OES and ICP-MS, and their contents increased with the prolongation of growth years. PCA results showed that Cu, Ni, Cd, Na, Mg, Fe, Ca, Zn, Mn and Hg were characteristic elements of Scutellariae Radix. Samples with different years could be divided into four categories in the spatial characteristic diagram of Fisher discriminant analysis. The correct rate of SVM model for identifying the growth years of samples was 95.2%. ConclusionThis established method is accurate and rapid for discriminating the growth years of Scutellariae Radix, which can provide reference for the identification of other Chinese medicinal materials. It is suggested that some elements should be considered as indexes in subsequent construction of the quality evaluation system of Scutellariae Radix.
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This paper aims to explore active components and mechanism of Scutellariae Radix(SR)-Phellodendri Chinensis Cortex(PCC) drug pair in treatment of psoriasis by network pharmacology and molecular docking. Specifically, the chemical components of SR and PCC were retrieved from literature and TCMSP, as well as targets of these components from PharmMapper and UniProt, and the targets related to psoriasis from OMIM, TTD, PharmGkb, and DrugBank. Then the chemical component-medicinal target, protein-protein interaction(PPI), and chemical component-psoriasis target networks were constructed by Cytoscape. Gene ontology(GO) term enrichment analysis and Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were performed based on Metascape. Finally, molecular docking of the chemical components(high degree) with core therapeutic targets was carried out by AutoDock vina. The results showed 88 compounds of SR and PCC(including baicalin, wogonoside, berberine and phellodendrine) and 30 targets of the pair in the treatment of psoriasis. The 30 targets mainly involved the biological processes such as neutrophil mediated immunity(GO: 0002446) and T cell activation(GO: 0042110), and the signaling pathways such as metabolism of xenobiotics by cytochrome P450(hsa00980), apoptosis(hsa04210), and PI3 K-Akt signaling pathway(hsa04151). The results of molecular docking demonstrated that the main active components can spontaneously bind to the targets and the binding energy of 46 components with epidermal growth factor receptor(EGFR) was less than-8 kcal·mol~(-1). According to the PPI analysis, EGFR may be a key target for the treatment of psoriasis. Active components such as baicalin and berberine had high binding affinity with EGFR. This study preliminarily revealed the multi-component, multi-target and multi-pathway mechanism of SR-PCC drug pair in the treatment of psoriasis, which provided theoretical basis for the research on the mechanism of the drug pair in the treatment of psoriasis.
Subject(s)
Drugs, Chinese Herbal , Molecular Docking Simulation , Psoriasis/genetics , Scutellaria baicalensis , Signal TransductionABSTRACT
Objective:To observe the effects of Scutellariae Radix-Hedyotidis Herba on the proliferation of human lung adenocarcinoma A549 cells and the expression of interleukin-6(IL-6), phosphatidylinositol 3-kinase (PI3K),protein kinase B (Akt), p-protein kinase B (p-Akt), mechanistic target of rapamycin (mTOR), hypoxia-inducible factor-1<italic>α </italic>(HIF-1<italic>α</italic>), and Cyclin D<sub>1</sub> at the cellular level, and to explore their molecular mechanism. Method:Following the set-up of the blank group (complete medium), low-, moderate-,and high-dose (20, 40, and 60 mg·L<sup>-1</sup>) Scutellariae Radix-Hedyotidis Herba groups, and low-, moderate-, and high-dose (5, 10, and 20 mg·L<sup>-1</sup>) cisplatin groups, the cell were treated with the corresponding drugs for 24, 48, and 72 h for detecting their viability by tetrazolium bromide (MTT) colorimetry. A549 cells were then divided into the blank group, Scutellariae Radix-Hedyotidis Herba group, cisplatin group, and combined medication group and intervened with the<sup> </sup>complete medium, 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba, 10 mg·L<sup>-1</sup> cisplatin, and 40 mg·L<sup>-1 </sup>Scutellariae Radix-Hedyotidis Herba + 10 mg·L<sup>-1 </sup>cisplatin, respectively, for 24, 48 and 72 h, followed by the measurement of inhibitory effects against the proliferation of A549 cells in each experimental group. The level of IL-6 in cell culture supernatant was determined by enzyme-linked immunosorbent assay (ELISA) after 72 h. The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in each group were assayed by real-time polymerase chain reaction (Real-time PCR), and the protein expression levels of PI3K, Akt, p-Akt, mTOR, HIF-1<italic>α</italic>, and Cyclin D<sub>1</sub> by Western blot. Result:After 24 h intervention, Scutellariae Radix-Hedyotidis Herba did not significantly inhibit the proliferation of A549 cells. However, 48 h later, the inhibitory effect in Scutellariae Radix-Hedyotidis Herba groups were significantly enhanced in comparison with that in the blank group (<italic>P</italic><0.05), exhibiting a time-dependent response. After 72 h of action, no significant change was present in the inhibitory effect of each Scutellariae Radix-Hedyotidis Herba group, so the optimal concentration of Scutellariae Radix-Hedyotidis Herba was set at 40 mg·L<sup>-1</sup> for follow-up experiments. As demonstrated by the comparison with the blank group, cisplatin at each concentration inhibited the cell proliferation in a time-dependent manner (<italic>P<</italic>0.05). Considering the cell survival rate, the best concentration of cisplatin was set at 10 mg·L<sup>-1</sup>. Compared with the blank group, Scutellariae Radix-Hedyotidis Herba combined with cisplatin remarkably inhibited the proliferation of A549 cells in a time-dependent manner (<italic>P<</italic>0.05), and the differences between the combined medication group and the other two groups became more significant after 72 h of medication (<italic>P<</italic>0.01). The IL-6 level in each experimental group, especially in the combined medication group, significantly declined in contrast to that in the blank group (<italic>P<</italic>0.01). The mRNA expression levels of HIF-1<italic>α</italic> and Cyclin D<sub>1</sub> in all experimental groups were obviously lower than those in the blank group, with the most significant changes observed in the combined medication group (<italic>P</italic><0.05,<italic>P</italic><0.01). The protein expression levels of PI3K, p-Akt, mTOR, HIF-1<italic>α</italic>, and Cyclin D<sub>1</sub> in each experimental group was significantly down-regulated(<italic>P</italic><0.05,<italic>P</italic><0.01), and the levels in the combined medication group were even lower than those in the cisplatin group (<italic>P<</italic>0.01). Conclusion:Scutellariae Radix-Hedyotidis Herba has an inhibitory effect on the proliferation of A549 cells, which may be related to its inhibition against the expression and secretion of IL-6/PI3K/Akt/mTOR-HIF-1<italic>α</italic> axis.
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Objective:To observe the difference in drying dampness between Scutellariae Radix and Atractylodis Rhizoma in model rats with spleen-stomach dampness-heat syndrome and clarify their property-efficacy relationship. Method:Sixty-four healthy male SD rats were randomized into the blank group, model group, high-, medium-, and low-dose Scutellariae Radix groups, as well as high-, medium-, and low-dose Atractylodis Rhizoma groups. The rats were exposed to high-fat and high-sugar diet and external dampness-heat environment for 20 days for inducing the spleen-stomach dampness-heat syndrome. The macroscopic manifestations of rats were observed and the morphological changes in stomach and colon were detected under a light microscope after hematoxylin-eosin (HE) staining, followed by the calculation of pathological scores. The serum tumor necrosis factor-<italic>α</italic> (TNF-<italic>α</italic>), interleukin-4 (IL-4), and interferon-<italic>γ</italic> (IFN-<italic>γ</italic>) levels were determined by enzyme-linked immunosorbent assay (ELISA). The protein and mRNA expression levels of aquaporin-4 (AQP4) in the gastric tissue were measured by Western blot and Real-time polymerase chain reaction (Real-time PCR), respectively. Result:Rats in the model group presented with the manifestations of dampness-heat syndrome. The inflammatory reaction in stomach and colon was obvious, and the pathological score was significantly increased (<italic>P</italic><0.01). The serum IFN-<italic>γ</italic>, IL-4, and TNF-<italic>α</italic> levels were elevated (<italic>P</italic><0.05), and so were the AQP4 protein and mRNA expression levels in the gastric tissue except that there was no statistical difference. The clinical symptoms of rats in the medication groups were alleviated. Scutellariae Radix significantly relieved the gastric and colonic inflammation in model rats. Atractylodis Rhizoma inhibited the colonic inflammation in model rats to a certain extent, but it had no obvious effect on gastric inflammation. The pathological score of each Scutellariae Radix group was decreased. In terms of the pathological score of gastric tissue, only the high-dose Scutellariae Radix produced a significant difference (<italic>P</italic><0.01), and the pathological scores of the three Atractylodis Rhizoma groups were not significantly different from that in the model group. As for the pathological score of colonic tissue, all the medication groups except for the low-dose Atractylodis Rhizoma group exhibited a significant difference in comparison with that of the model group (<italic>P</italic><0.01). Scutellariae Radix and Atractylodis Rhizoma at each dose reduced not only the serum IFN-<italic>γ</italic>, IL-4, and TNF-<italic>α</italic> levels (<italic>P</italic><0.05, <italic>P</italic><0.01), but also the AQP4 protein expression in gastric tissue of model rats (<italic>P</italic><0.01). The AQP4 mRNA expression in the gastric tissue of model rats declined in the high- and low-dose Scutellariae Radix groups, while that in the medium-dose Scutellariae Radix group and each Atractylodis Rhizoma group rose without statistical difference. Conclusion:Scutellariae Radix and Atractylodis Rhizoma display a certain property-efficacy relationship in drying dampness of rats with spleen-stomach dampness-heat syndrome. Specifically, the efficacy of drying dampness is related to their cold/heat property, and the resulting outcome of bitter-cold Scutellariae Radix is better than that of bitter-warm Atractylodis Rhizoma.
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In the pharmacopoeia, many process parameters for the purification process of Scutellariae Radix are unclear. In this study, deterministic screening design combined with design space method was used to optimize the purification process of Scutellariae Radix extract. Nine method parameters such as mass fraction of solution(X_1), first acid precipitation pH(X_2) and first holding time(X_3) in the purification process were firstly studied by definitive screening design. The yield of baicalin was defined as the evaluation index. A stepwise regression method was used then to build quantitative models between evaluation index and method parameters and the three most critical impact parameters were determined. Probability-based design space was calculated and successfully verified with the experimental error simulation method. Finally, the second standing temperature, the first standing temperature and the pH value of the second acid precipitation were determined as the three most critical method parameters. The recommended operating space was as follows: the second standing temperature 5-7 ℃, the first standing temperature 13-15 ℃, and the pH of the second acid precipitation 1.5-1.7. Within this operating space, the baicalin yield in the purification process was over 80%, and the probability of reaching the standard was over 0.96. In this study, we optimized the effect of various parameters for the purification process of the Scutellariae Radix extract in the pharmacopoeia on the yield of baicalin and provided a reference for industrial production of the exact of Scutellariae Radix.
Subject(s)
Drugs, Chinese Herbal , Flavonoids , Plant Extracts , Scutellaria baicalensisABSTRACT
Objective::To compare the pharmacodynamic effects of pith-nodecayed and pith-decayed products of Scutellariae Radix on rats with large intestine damp-heat syndrome, and to demonstrate the scientificness of dividing Scutellariae Radix into pith-nodecayed and pith-decayed products as medicines by modern pharmacological test. Method::Rats were randomly divided into blank group, model group, low-and high-dose group of pith-nodecayed products (0.9, 3.6 g·kg-1), low-and high-dose group of pith-decayed products (0.9, 3.6 g·kg-1), Scutellariae Radix group (0.9 g·kg-1), compound berberine tablets group (positive drug group, 0.045 g·kg-1), and 8 rats in each group. Taking model rats with large intestine damp-heat syndrome, the body temperature, thymus index, spleen index, pathological sections of colon and ileum, inflammatory factors and Secretory immunoglobulin (SIg) A content were selected as indexes to evaluate the therapeutic effect of pith-nodecayed and pith-decayed products on large intestine damp-heat syndrome, and make comprehensive evaluation of the difference in efficacy between them. Partial least squares-discriminant analysis (PLS-DA) was employed to analyze the pharmacological indexes of these two products against large intestine damp-heat syndrome. Result::Pith-nodecayed and pith-decayed products of Scutellariae Radix with different doses could reduce the body temperature, thymus index, spleen index, contents of interleukin (IL)-2, IL-6, IL-1β in serum and SIgA content in intestinal mucosa, and most of them had significant differences (P<0.05, P<0.01). Compared with the isodose group of pith-decayed products, the effect of corresponding dose group of pith-nodecayed products was better, and most of them had significant differences (P<0.05, P<0.01). PLS-DA results indicated that there were significant differences in the pharmacological effects of pith-nodecayed and pith-decayed products, and they were clustered on one side, respectively. Conclusion::Both of pith-nodecayed and pith-decayed products of Scutellariae Radix have therapeutic effect on large intestine damp-heat syndrome with distinctly different strength of action, and pith-nodecayed products is superior to pith-decayed products, which verify the scientific nature of pith-nodecayed products was specializedly used to treat bowel disease in ancient times.
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Objective:The aim of this study was to research the relationship between HPLC fingerprint and anti-inflammatory effect of Zhideke granules, and the substance basis of its anti-inflammatory effect was preliminary explored. Method:The fingerprint of 10 batches of Zhideke granules were determined by HPLC, the mobile phase was consisted of methanol-0.2% phosphoric acid solution for gradient elution with a detection wavelength of 254 nm. Similarity analysis, hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) were used to evaluate the quality difference between batches of Zhideke granules. The correlation analysis between the common peaks and the inhibition rates of Zhideke granules on ear swelling and cotton ball granuloma in mice was carried out by partial least squares (PLS), and the peaks greatly contributing to the anti-inflammatory effect were screened out. Result:There were 31 common peaks in the HPLC fingerprint of Zhideke granules. The similarities of 10 batches samples were ≥0.992. The HCA and PCA analysis results were consistent, and the samples were divided into 3 categories. Combined with the OPLS-DA results, 15 components were the main markers affecting the differences of different batches of samples. Different batches of Zhideke granules differed in anti-inflammatory effect. The chromatographic peaks being positively correlated with the anti-inflammatory effect were mainly from Belamcandae Rhizoma and Scutellariae Radix, Chromatographic peaks 3, 6, 19, 27-30 had significant contribution to anti-inflammatory effect, of which peaks 28 and 30 were irisflorentin and wogonin. Conclusion:HPLC fingerprint combined with chemical pattern recognition method can provide a reference for systematic evaluation of the overall quality of Zhideke granules. Zhideke granules has a certain inhibitory effect on acute and chronic inflammation in mice, and the anti-inflammatory effect is the result of the combined action of various ingredients, while Belamcandae Rhizoma and Scutellariae Radix have significant significance for the anti-inflammatory effect.
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Objective: HPLC fingerprints of Banxia Xiexin Decoction (BXD) was established to analyze the quality value transmitting relationship between medical herbs-decoction pieces-whole decoction of Scutellariae Radix (SR) and Coptidis Rhizoma (CR). Methods: BXD was prepared by the decoction method recorded in Treatise on Febrile Diseases. BXD HPLC fingerprints was established, and the characteristic peaks were analyzed; The transfer rate, the dry extract rate and the similarity of fingerprint were used as indicators to analyze the quality value transmitting relationship between medical herbs-decoction pieces-whole decoction of SR and CR. Results: The transfer rate of baicalin from medical herbs to decoction pieces was between 81.08% and 119.82%; The transfer rate of baicalin from decoction pieces to BXD was between 44.55% and 59.05%; The transfer rate of berberine from medical herbs to decoction pieces was between 66.65% and 97.51%; The transfer rate of berberine from decoction pieces to BXD was between 7.83% and 22.93%; And the dry extract rate of the whole decoction was between 12.99% and 17.21%. The similarity of fingerprints of 15 batches of BXD ranged from 0.978 to 0.999. Conclusion: The quality evaluation method established by the fingerprint is applicable to the quality control of medical herbs-decoction pieces-whole decoction of SR and CR, which provides some reference for the establishment of the quality evaluation method of famous classic formula.
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Objective: To establish the quality evaluation system based on spectrum-antibacterial effect correlation of Yinhuang Granules in order to detect its raw medicinal materials, extracts, and preparation simultaneously. Methods: Firstly, the spectrum- antibacterial effect correlation quality evaluation system of Yinhuang Granules was established by using Least Squares Support Vector Machine (LS-SVM). Then, by using supervised partial least square-discriminant analysis models (PLS-DA), the antibacterial effect evaluation was judged based on the spectrum-antibacterial effect data of Scutellariae Radix, Lonicerae Japonicae Flos, Scutellaria extract, Lonicerae japonica extract, and Yinhuang Granules. Results: The mathematical model of Yinhuang Granules based on spectrum-antibacterial effect correlation was established; The average relative error of the prediction results was less than 5%, and the antibacterial rate of Scutellariae Radix, Scutellaria Radix extract, Lonicerae Japonicae Flos, Lonicerae japonica extract was greater than 43%, 5.5%, 11%, 37% calculated by their mathematical model, which can ensure the antibacterial effect was greater than 11% (correct rate was 87%). Conclusion: The quality evaluation system can realize the quality control of the key link of the production of traditional Chinese medicine, the evaluation results are more scientific, comprehensive, and accurate.
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OBJECTIVE: To develop a method for determining 12 constituents in Yigan Jiedu Capsules by HPLC. METHODS: The quantitative analysis was carried out on a Waters Atlantic T3 C18column maintained at 40 ℃, using mobile phase consisting of acetonitrile(A)and 0.1% phosphoric acid solution(B)by a gradient elution program(0-10 min, 4%→13%A; 10-20 min, 13%→15%A; 20-25 min, 15%→19%A; 25-35 min, 19%A; 35-36 min, 19%→23%A; 36-45 min, 23%→27%A; 45-56 min, 27%→43%A; 56-65 min, 43%→65%A) at a flow rate of 1 mL•min-1. The detection wavelength was set at 270 nm in the first 22 min, and then changed to 290 nm between 22 and 44 min, and last changed to 270 nm. RESULTS: The linear ranges of gallic acid, vanillic acid, picroside Ⅱ, astilbin, engeletin, picroside, baicalin, berberine hydrochloride, wogonoside, emodin, chrysophanol and wogonin fell with in the range of 0.012 19-0.243 8 μg, 0.036 70-0.734 0 μg, 0.033 77-0.675 4 μg, 0.057 38-1.147 6 μg, 0.006 580-0.131 6 μg, 0.015 38-0.307 6 μg, 0.067 11-1.342 2 μg, 0.028 04-0.560 8 μg, 0.041 48-0.829 6 μg, 0.006 310-0.126 2 μg, 0.013 64-0.272 8 μg and 0.014 22-0.284 4 μg. The recoveries were 98.7%, 98.9%, 99.4%, 99.2%, 98.9%, 99.1%, 98.9%, 99.6%, 99.3%, 98.9%, 98.6% and 99.0%, respectively. The relative standard deviations were 1.02%, 0.46%, 0.64%, 0.86%, 1.1%, 1.3%, 0.60%, 0.92%, 0.61%, 1.5%, 0.93%and 0.98%, respectively(n=6). CONCLUSION: This method is simple, accurate, reproducible and convenient for the quality control over Yigan Jiedu Capsules.